Fig 2: sOPN was increased expressed in OA subchondral bone mainly in preosteoblasts and osteoblasts. (A, B) Western blot and qPCR analysis of the expression of OPN in primary osteoblasts of subchondral bone of an OA mouse model and control group; n = 3. (C, D) Representative immunofluorescent and quantitative analysis of sOPN in OSX+ preosteoblasts or OCN+ osteoblasts in the tibial subchondral bone of an OA mouse model and control group. Positive cells were indicated with arrows. The boxed area is magnified in the corner, scale bars = 50 µm. Data are shown as mean ± s. d. and were analyzed by Student’s t test; n = 6, *P < 0.05,**P < 0.01.

Fig 3: Detection of OPN expression levels in cancer tissues and para-carcinoma tissues via H&E and IHC. Cancer and para-carcinoma tissues of the same patient are shown (A-D). (A and B) Clear cell adenocarcinoma is shown in H&E staining, and cells in the lesion region display the cystic, papillary and hyaline changes. (C) The negative expression of OPN. (D) OPN is located in the cytoplasm, and the expression is strongly positive. OPN, osteopontin.

Fig 4: Effect of lentivirus interference in OPN on cell proliferation capacity. OPN, osteopontin. *P<0.05; **P<0.01.

Fig 5: Peripheral biliary differentiation is dependent on both Notch signaling and substrate stiffness.(A) Immunolabeling for OPN of BMEL cells presented with DLL4 on 30 kPa and 4 kPa substrates. Cells were treated with vehicle control (DMSO) or an inhibitor of Notch signaling (?-secretase inhibitor X, GSI, 5 µM). (B) Quantification of OPN+ cell counts on 30 kPa and 4 kPa substrates after treatment with DMSO or GSI. (C) Immunolabeling for SOX9 and HNF4A of BMEL cells on 30 kPa and 4 kPa substrates. (D) Quantification of SOX9 and HNF4A intensity on 30 kPa and 4 kPa substrates. (E) RNA in situ hybridization for Jag1, Dll1, and Notch2 on 30 kPa and 4 kPa substrates. Cells were exogenously presented with IgG or DLL4. (A, C, E) Scale bars indicate 150 µm. (B, D) Mean ± 95% CI.10.7554/eLife.38536.013Figure 2—source data 1.Summary table for OPN data in Figure 2B and Figure 2—figure supplement 1.10.7554/eLife.38536.014Figure 2—source data 2.Summary table for SOX9 and HNF4A data in Figure 2D.